New steroid alaninates



United States Patent 2,979,518 NEW STEROID ALANINATES 6 David Adriaan van Dorp and Hendrik Paul de Jongh,

Oss, Netherlands, assignors to Organon Inc., Orange,

As starting substances for the preparation of these newamino acid esters are employed steroids which have one or more esterifiable hydroxyl groups and which contain at least one double bond in the nucleus and which may in addition be substituted by one or more free or functionally converted hydroxyl and oxo groups, halogen atoms and saturated or unsaturated lower hydrocarbon groups.

By a functionally converted hydroxyl group is understood an etherified or esterified hydroxyl group. The etherified hydroxyl group may e.g. be a methoxy, tetrahydropyranyloxy, benzyloxy, or tn'phenylrnethoxy group. An esterified hydroxyl group may be a hydroxyl group esterified with an aliphatic, aromatic or araliphatic carboxylic acid, e.g. formic acid, acetic acid, cyclohexyl butyric acid, benzoic acid, and p-phenylpropionic acid.

By a functionally converted 0x0 group is understood e.g. an enolester, enolether, acetal, mercaptal, enamine, oxim and hydrazone grouping.

The free or functionally converted hydroxyl and 0x0 groups possibly present in the starting substances may occur e.g. in the positions 1, 2, 3, 4, 6, 7, 8, 9, 11, 12, 14, 15, 16, 17, 20, and 21.

A lower hydrocarbon group which may be both saturated and unsaturated, e.g. a methyl, ethyl, vinyl, and ethinyl group, may occur e.g. in the positions 1, 2, 4, 6,

l0, l3, and 17-.

A halogen atom may e.g. occur in the positions 1, 2, 4, 6, 9, 11, 12, 15, 16, and 21.

The hydroxy steroids to be applied as starting substances may have any steric configuration and may be 18-nor, 19-nor, 18,19-bisnor, C-nor or D-horno steroids. Especially the unsaturated hydroxy steroids of the androstane, oestrane, and pregnane series are employed for the present esterification.

The esterification may take place at one or more of the esterifiable hydroxyl groups. Especially the hydroxyl groups, present in the positions 3,-11, 16, 17, and 21, are considered for this purpose.

The following steroids are mentioned by name as starting substances for the present process:

A -17p-hydroxy-3-oxo-androstene A -17p-hydroxy-3-oxo-17a-methyl androstene A -17p-hydroxy-3-oxo-17a-vinyl-androstene A -17phydroxy-3-oxo 19-nor-androstene A -3p,17B-dihydroxy-androstene A -3p-hydroxy-17-oxo-androstene 2,979,518 Patented Apr. 11, 1961 A -3-hydroxy-17-oxo-oestratriene A -3,17p-dihydroxy-oestratriene A -3,17p-dihydroxy-l7a-ethinyl-oestratriene A -3,17p-dihydroxy-1-methyl oestratriene A -3,16u,17p-trihydroxy-oestratriene A -17p-hydroxy-S-oXo-androstadiene A -21-hydroxy-3,20-dioxo-pregnene A -1 1,6,21-dihydroxy-3,20-dioxo-pregnene A -2 1-hydroxy-3 ,'l 1 ,20-trioxo-pregnene A -17a,21-dihydroxy-3,20-dioxo-pregnene AM 1 p,1701,21-trihydroxy-3,ZO-dioxo-pregnene A -17a,2l-dihydroxy-3,1 1,20-trioxo-9a-fluoro-pregnene A -17u,21-dihydroxy-3,1 1,20-trioxo-pregnene A -17a-hydroxy-3,20-dioxo-pregnene A-20-oxo-3B-hydroxy-pregnene A -17a,21-dihydroxy-3,11,20-trioxo-pregnadiene A -1 1 8,17 11,2 l-trihydroxy-B,ZO-diOXo-pregnadiene A 17a,21-dihydroxy-3,l 1,20-trioxo 9 t-fiuoro-pregnadiene A -1 1 p, 17 (1,21-trihydroxy-3,20-dioxo- 9u-fluoro-preguadiene Especially the N -oestratriene compounds and the A and A -3-oxo compounds of the androstane and pregnane series, which have an activity in the non-esterified form, result, in the present esterification, in important biologically active compounds.

The esterification of the unsaturated hydroxy'steroids is carried out with an aminomonocarboxylic acid which can be employed in one of its optically active forms or as a racemate. The amino acids may be e.g. monoaminomonocarboxylic acids and diaminomonocarboxylic acids, such as glycine, aand [Si-alanine, valine, leucine, norleucine, isoleucine, norvaline, a-amino btuyric acid, phenylalanine, proline, lysine, ornithine, and amino benzoic acid, such as p-arnino benzoic acid.

For the esterification of the hydroxy steroids with amino acids according to the invention it is necessary that the amino group(s) of the latter compounds be protected. As a protecting group there is preferably employed a carbobenzoxy group, such as has been described in Advances in Protein Chemistry, vol. V, page 25 and following pages.

The esterification of the hydroxy steroid with e.g. a carbobenzoxy amino acid may take place by carrying out the reaction in the presence of a dehydrating agent, e.g. ethoxyacetylene. The ester in question can also be prepared by reaction of the anhydride or acid chloride of the used amino acid derivative with the hydroxy steroid in question.

After the esteterification the carbobenzoxy group can be split ofi e.g. by catalytic reduction as has been described in Advances in Protein Chemistry, vol. V, page 29 and following pages. The protecting group, however, is preferably split off by means of a hydrohalogenic acid, e.g. hydrobromic acid. The advantage of this method is that the double bonds present in the esterified steroid are not reduced.

The esterification of the hydroxy steroid with a carbobenzoxy-aminomonocarboxylic acid is e.g. carried out by heating the two components in a suitable solvent and in the presence of a dehydrating agent for some time. As a solvent may be used e.g. an ether, such as methyl ethyl ether, diethyl ether, dioxane and tetrahydrofurane, an ester, such as ethyl acetate, an aromatic hydrocarbon, e.g. benzene, a ketone, such as acetone, a halogenated hydrocarbon, e.g. dichloroethane, or a mixture of these solvents. x

Usually the reaction time lies between 1 and 24 hours, while the reaction temperature amounts to about 3 It is also possible to carry out the esterification by In case it is desired to split off the carbobenzoxy groupafter the .esterification--by means of a catalytic reduction, the o e-unsaturated oxo grouping possibly present in the steroid compound should be protected e.g. by ketalization. This protection may also be carried out already prior to the esterification. From the formed ester the carbobenzoxy group is now split off in a neutral medium by catalytic reduction in a suitable solvent, such as an alcohol, an ether, an ester, or a mixture thereof. After filtering ofi the catalyst and evaporating the solvent in vacuo, the ketalized amino acid steroid ester is introduced into water and stirred in it for some hours in the presence of a little inorganic acid. As an acid there may be used hydrochloric acid, hydrobromic acid, or hydriodic acid. The reaction time lies between about 1 and 8 hours and the reaction temperature between about 15 and 80 C. In this manner the liberation of the nap-unsaturated ketone and the formation of the halogen salt of the new ester are simultaneously efiected. After filtration or centrifugation the ester is taken up in water, after which the aqueous solution is evaporated to dryness in vacuo or lyophilized. In this manner the salt of the amino acid ester is obtained in a dry form.

The protecting earbobenzoxy group is preferably split off by means of a hydrohalogenic acid, e.g. hydrobromic acid. This splitting is carried out e.g. in absolute dioxane or glacial acetic acid. In this manner the salt of the applied-hydrohalogenic acid, e.g. the HBr-salt, of the amino acid steroid esters is obtained directly.

of some compounds, prepared according to the present process, especially the salts, it was found that they exert a favourable action on the fat metabolism. In addition these compounds are of importance owing to the antiphlogistic action which some of these compounds show on local application.

Example Ia 9.96 g. of A -3-ethylenedioxyandrostene-17,9-01, 6.72 g. of N. carbobenzoxyglyeine, 3 ml. of ethoxyacetylene in 150 ml. of benzene are heated under gentle boiling for 5 hours. During the reaction everything goes into solution. After completion of the reaction the reaction mixture is evaporated to dryness in vacuo. The residue is taken up in little benzene. Added to this are about 500 ml. ethanol with 1 drop of pyridine. Evaporation to about 200 ml. at normal pressure. After cooling crude A -3-ethylene-dioxyandrostene-175-01-17 N. carbobenzoxy glycinate crystallizes out. Yield 8 g. or 51% of the theoretical. M.P.: 160-165 C. Recrystallization from ethanol with 1 drop of pyridine yields the pure compound of M.P. l66-l69 C. (a) =-33.6 (chloroform).

Analysis.-Oalculated for C H O N: N=2.67%.

Example 1b 6 g. of A -3-ethylenedioxy-androstene-173-01-17-N. carbobenzoxyglycinate are dissolved, while heating, in 600 ml. of 96% ethyl alcohol. After cooling 3 g. of palladium on carbon (5% palladium) are added and hydrogen is led through for 3 hours with violent stirring at a temperature of about 30 C. The escaping gas mixture is led through barium hydroxide (saturated solution) and the formed barium carbonate is weighed. After completion of the reaction the solution is filtered over hyflo from the catalyst. The residue on the filter is washed with some ml. of chloroform. The filtrate is evaporated to dryness in vacuo. The crude compound is dissolved in about 200 ml. of benzene and filtered off from the insoluble fraction. The filtrate is evaporated to dryness in vacuo. The yield of A -3-ethylene-dioxyandrostene-17p-ol-17-glycinate is 3.6 g. or 81% of the 4 theoretical. M.P.: 186-190 C. (decomposition). To obtain pure ethyleneketal of testosterone glycinate the crude productis recrystallized from benzene-petroleumether (60-80 C.), 2:1. M.P.: 195-199 C. (decomposition). (a) =-48 (chloroform).

Analysis.Calculated for C H O N: N=3.59%. Found: N=3.56%.

Example 10 Suspend 1.28 g. of A -3-ethylenedi0xy-androstene-17B- ol-l7-glycinate in ml. of H 0 and 2 ml. of 2 N HCl. Stir at room temperature for 3 hours. Centrifuge the cloudy solution. Decant and lyophilize the, clear aqueous solution. Yield: 0.76 g. or 60.5% of the theoretical of hydrochloric acid salt of 3-keto-A -androstene-173-01-17- glycinate. Decomposition at about 200 C. U.V. absorption:

Lambda max. (in alcohol): 24111111. Epsilon: 14,100 (in alcohol) Lambda max. (in water): 248 mp Epsilon= 13,500 (in water) (a) =+67.9 (ethanol) Analysis-Calculated for C H O NCl: N=3.66%.

Found: N=3.33%.

Example Ila 7.2 g. of testosterone, 5.5 g. of carbobenzoxyglycine, and 2.6 ml. of ethoxyacetylene are dissolved in ml. of benzene. Heating, while stirring, under gentle boiling for 6 hours. Evaporation in vacuo and taking up the residue in ether after completion of the reaction. The 3-ketone-A -androstene-l7B-ol-N. carbobenzoxyglycinate crystallizes out slowly. After filtering ofi the substance is recrystallized from methanol. M.P.: 162-1635 C. (a) =+69.6 (chloroform).

Analysis.Calculated for C H O N: N =2.92%. Found: N=3.03%.

From the testosterone have been prepared in a corresponding manner: A -3-oxo-17fl-hydroxy-androstene- 17fl-(N-carbobenzoxy)-alaninate and A -3-oxo-17p-hydroxy-androstene-l7fi-(N-carbobenzoxy)-phenylalaninate.

Example 11b 5 g. of 3-keto-A -androstene-l7p-ol-N. carbobenzoxyglycinate are dissolved in ml. of absolute dioxane. Now, for 100 minutes, dry HBr-gas is led through while coohng with ice water. The 3-keto-A -androstene--01- l7-glycinate-HBr separates as a reddish-brown oil. Evaporation in vacuo as a result of which the oil crystallizes. The crystals are dissolved in water, the solution is shaken out with petroleum-ether (40-60) to remove formed benzylbromide, treated with norit, filtered, and lyophilized. Yield quantitative. Decomposition above 270 C. (a) =-|-64.8

(ethanol). U.V.-absorption:

Lambda max. (alc.)=24l mp Epsilon= 14,400

Analysis-Calculated for CgjHaaOgNBl'i N=3.28%. Found: N=3.15%.

In an analogous manner the following water soluble salts have been prepared: A -3-oxo-17p-hydroxy-androstene-l7p-alaninate-HBr and A 3 oxo 175 4 hydroxyandrostene-l7p-phenylalaninate-HBr.

Example Illa A solution of 21.76 g. of A -3,17p-dihydroxy-oestratriene (oestradiol) and 33.17 g. of carbobenzoxy-glycine in 700 ml. of 1,2-dichloroethane and 17 ml. of ethoxyacetylene is refluxed, with stirring, for 16 hours. After completion of the reaction the reaction mixture is evaporated to dryness in vacuo. The residue is chromatographed over silica gel, in which benzene with an increasing acetone content is used as elution agent. From the benzene-acetone fractions (95:5) 21.75 g. of oestradiol-3,17p-di-(N-carbobenzoxy) -glycinate are'obtained as an oil; (a) =+21.0 (chloroform). k

Analysis-Calculated for c mgomg N=4.28%.

- been prepared:

Oestradiol-3,17fl-di(N-carbobenzxy)-alaninate Oestradiol-3,17B-di(N-carbobenzoxy)-phenyl alaninate Example lllb To a solution of 13.1 g. of oestradiol-3,17p-di-,(N-'carbobenzoxy)-glycinate in 75 ml. of absolute dioxane are added 42.5 g. of a HBr solution in dioxane containing 19% by weight of HBr. The mixture is shaken at room temperature for 90 minutes, after which the'formed precipitate is filtered off. This precipitate is dissolved in water, after which the solution is treated with nori The thus obtained clear aqueous solution is lyophilized, after which 5.72 g. of crude oestradiol-3,17B-diglycinate-HBr are obtained. Recrystallization from a mixture of ethanol and ether yields white crystals which decompose at 245- 255 C.; (a) =+23.6 (ethanol). 1

Analysis.Calculated for C H ,O N,Br,: Found: N=5.10%. Found: N=4.88%.

In a corresponding manner the following estershave been prepared:

Oestradiol-B,l7fl-di-alaninate-Hl3r Oes tradiol-3,17B-di-phenyl alaninate-HBr Example I Va 1 18 g. of A -3,11,20-trioxo-l7a,2l-dihydroxy-pregnene, 10.75 g. of carbobenzoxyglycine in 250ml. of 1,2-dichloroethane and 6 ml. of ethoxyacetylene are heated under reflux, while stirring, for 6.5 hours. The resulting clear solution is evaporated to dryness in vacuo. The residue is taken up in 250 ml. of ethanol, after which this solution is evaporated to about 150 ml. After48 hours at room temperature the A -3,1l,20-trioxo-l7a,21-dihydroxy-pregnene-2l-(N-carbobenzoxy) glycinate crystallizes out. The M.P. hereof is 186.5-187.5' C.; (a) =-|-171.5 (chloroform). Lambda max (ethanol)=238 ma; epsilon=15,100.

Analysis.-Calculated for C H O N: Found: N=2.45%.

In a corresponding manner the following N-carbobenzoxy-amino acid esters have been prepared:

Cortisone-2l-(N-carbobenzoxy)-alaninate and Cortisone-2 l- (N-carbob enzoxy) -phenyl alaninate Example I Vb 5.5 g. of A -3,11,20-trioxo-l7a,2l-dihydroxy-pregnene- 2l-(N-carbobenzoxy)-glycinate are dissolved in ml. of absolute dioxane. Then 6 ml. of dioxane containing 24.2% hydrobromic acid are added to the solution. The reaction mixture is kept at room temperature for 90 minutes, after which the dioxane is evaporated in vacuo. The oily residue is taken up in water and then extracted with chloroform. The aqueous layer is separated, treated with norit, filtered and then lyophilized.

After recrystallizing the' resulting pale yellow powder from a mixture of absolute and ethanol and ether the A4-3,1 l ,20-t1'iOXO-l7a,2 l-dihydroxy-pregnene-2 l-glycinate- HBr is obtained. The compound melts under decomposition. (a) =+164 (ethanol).

Analysis-Calculated for C H O N Br: N=' Found: N=2.58%.

Lambda max. (alc.)=238 mp; epsilon=l4,500 Lambda max. (water) =245 mp; epsilon= 15,400

After acetylaticn the A*-3,l1,20-trioxo-17a,21-dihydr xy-preg'nene-Zl-(N-acetyl)-glycinate is obtained; M.P.

Analysis.,Calculated for C H O- N: N=3.05%.

- Found: N=3.34%.

' In an almost analogous manner have been prepared:

Cortisone-2l alaninate-HBr and Cortisone-Zl-phenyl alaninate-HBr Example Va in:250 ml. of absolute -dioxane and 6 ml. of ethoxy acetylene is heated at 90 C. for 7 hours. Subsequently the reaction mixture is evaporated to dryness in vacuo. The. residue is dissolved in 300 ml. of ethanol, after which this solution is evaporated to half the volume. A-fter standing for some time the A -3,l1,20-trioxo-17,2l-dihydroxy-pregnadiene 21 (N-carbobenzoxy)-glycinate crystallizes out. The M.P. hereof is 186- 189 C. Yield: 14.91 g. After recrystallization from ethanol the pure compound of M.P. 188-489 C. is obtained; (a) =+152.5 (chloroform).

Analysis.-Ca1culated for C H O;N: N=2.55%. Found: N=2.40%.

Lambda max. (alc.)=239 mp Epsilon: 15,000

In an analogous manner the following esters of prednisone have been prepared:

Prednisone-Zl-(N-carbobenzoxy) -alaninate Prednisone-21-(N-carbobenzoxy) -phenyl alaninate Example Vb Lambda max. (ethanol) =238 m epsilon= 14,400 Lambda max. (water)=244 m epsilon=l4,400

Analysis.-Calculated for c H o NBr: N=2.80%. Found: N=2.56%.

I Ina corresponding manner the following salts have been prepared:

Prednisone-Zl-alaninate-HBr Prednisone-Zl-phenyl alaninate-HBr Example Vla 21.6 g. of A -3,20-dioxo-1lp,7a,2l-trihydroxy-pregnadiene and 15 g. of carbobenzoxy-d-alanine are dissolved in 250 ml. of absolute dioxane. 6 m1. of ethoxyacetylene are added to this solution, after which the mixture is further treated in an analogous manner as described in Example Va. Obtained are 15.5 g. of prednisolone-2l-(N-carbobenzoxy)-d-alaninate containing 1 mol of crystal alcohol. To remove the crystal alcohol the compound is dissolved in chloroform and precipitated therefrom with ether. The M.P. is l56157.5 C.; (a) ='-|-103.7' (chloroform). Lambda max. (ethanol)=243 m epsilon= 14,800.

Analysis.Calculated for C H O N: N=2.47%. Found: N=2.39%.

In an analogous manner the following derivatives of the prednisolone have been prepared:

Prednisolone-21-(N-carbobenzoxy)-glycinate and PredmsoIone-ZI-(N-carwbemxy)-phenyl alaninate l the the following salts .Found: N=2.45%

7- Example VIb 13.55 g. of A -3,20-dioxo-11p,17u,21-tiihydroxy-pregnadiene-2l-(N-carbobenzoxy)-d-alaninate are dissolved in 60 ml. of absolute dioxane. To this solution are added ml. of absolute dioxane containing 0.49 g. of HBr per ml. of dioxane. The reaction mixture is shaken at room temperature for 90 minutes, in which an oily residue is formed. The mixture is then evaporated to dryness in vacuo, after which the residue is dissolved in water. This aqueous solution is washed with ether, treated with norit, filtered, and then lyophilized. Obtained are 9.5 g. of prednisolone-2l-d-alaninate-HBr. After a few recrystallizations from a mixture of absolute ethanol and ether the pure compound is obtained; (a) =+l06 (96% ethanol).

Lambda max. (ethanol)=243mp; epsilon=14,100 Lambda max. (water)=2 47 m epsilon=13,700

Analysis-Calculated for C H O NBr: N=2.73% Found: N=2.45%.

In an analogous manner the following amino acid esters have been prepared:

Prednisolone-Zl-glycinate-HBr and PrednisoloneQl-phenyl alaninate-HBr Example VIIa 10.86 g. of A -3-20-dioxo-1lfl,17a,21-trihydroxy-pregnene, 6.40 g. of carbobenzoxyglycine in 130 ml. of dichloroethane, and 3 ml. of ethoxyacetylene are heated for 8 hours, while stirring. Then the reaction mixture is evaporated to dryness in vacuo and the residue is subsequently treated in an analogous manner as described in Example IVa.

Obtained is the A-3,20-dioxo-11p,17u,21-trihydroxypregnene-2 1- (N-carbobenzoxy) -glycinate.

Analysis.-Calculated for C H O N: N=2.53%. Lambda max. (alc.)=242 mp; epsilon= 16,000.

In a corresponding manner the following N-carbobenzoxy-amino acid esters of hydrocortisone have been prepared:

Hydrocortisone-21-(N-carbobenzoxy)-alaninate Hydrocortisone-2 l (N-carbobenzoxy) -phenyl alaninate Example VlIb epsilon: 14,500.

In an analogous manner the following salts have been prepared:

Hydrocortisone-2i-alaninate-HBr Hydrocortisone-2l-phenyl alaninate-HBr Example VIIIa 16.44 g. of 19-nor-testosterone and 1 g. of carbobenzoxydl-phenyl alanine are suspended in 1 ml. of benzene. 7 ml. of ethoxy acetylane are added to this solution, after which the reaction mixture is refluxed for 8 hours. After completion of the reaction the mixture is evaporated to dryness in vacuo, after which the residue is chromatographed over silica gel. As elution agent is applied benzene with an increasing ether content. From .the benzene-ether fractious (23:2) 19.63'g. of 19-nor testosterone-17fl-(N-carbobenzoxy)-dl-phenyl alaninate are isolated. The M.P. is 55-60 C.; (a) =+3l.7 C. (chloroform).

Analysis-Calculated for C H O N: N=2.52%. Found: N=2.63%.

Lambda max. (96% ethanol) =240 m Epsilon=1'6,100

In a corresponding manner the following derivatives have been prepared:

l9-nor-testosterone-17fi- (N-carbobenzoxy) -glycinate and 19-nor-testosterone-171S-(N-carbobenzoxy) -alaninate Example VIIIb 4 g. of 19-nor-testosterone-17,3(carbobenzoxy)-dl-phenyl alaninate are dissolved in ml. of absolute dioxane. In an analogous manner as described in Example IIIb the mixture is converted into the 19-nor-testosterone-l7fi-dlphenyl alaninate-HBr.

Analysis.-Calculated for C, H O NBr: N=2.79%. Found: N=2.51%

Lambda max. (alc.)= 239 III/1.; epsilon=15,000.

In the same manner the following salts have been prepared:

19-nortestosterone-17,6-glycinate-HBr and l9-nor-testosterone-17p-alaninate-HBr We claim:

1. A compound selected from the group consisting of oestradiol-3,17-di-alaninate and the acid addition salts thereof.

' 2. A compound selected from the group consisting of oestradiol-3,l7-di-phenylalaninate and the acid addition salts thereof.

3. A compound selected from the group consisting of cortisone-Zl-alaninate and the acid addition salts thereof.

4. A compound selected from the group consisting of cortisone-Zl-phenylalaninate and the acid addition salts thereof.

5. A compound selected from the group consisting of prednisone-Zl-alaninate and the acid addition salts thereof.

6. A compound selected from the group consisting of prednisone-2l-phenylalaninate and the acid addition salts thereof.

7. A compound selected from the group consisting of prednisolone-2l-alaninate and the acid addition salts thereof.

8. A compound selected from the group consisting of prednisolone-Zl phenylalaninate and the acid addition salts thereof.

9. A compound selected from the group consisting of hydrocortisone-Zl-alaninate and the acid addition salts thereof.

'10. A compound selected from the group consisting of hydrocortisone-2l-phenylalaninate and the acid addition salts thereof.

References Cited in the file of this patent UNITED STATES PATENTS 2,173,423 Miescher et a1 Sept. 19, 1939 2,547,949 Lawson et al Apr. 10, 1951 2,547,961 Mooradian et a1 Apr. 10, 1951 2,660,586 Murray et -al Nov. 24, 1953 2,708,651 Laubach May 17, 1955 2,751,402 Schneider June 19, 1956 2,816,902 Gould et a1 Dec. 17, 1957 2,818,408 Campbell et al Dec. 31, 1957 2,838,534 Babcock et al June 10, 1958 2,840,581 Hogg et al. June 24, 1958 2,871,160 Johnson et al Jan. 27, 1959 2,885,413 Hogg et a1 May 5, 1959 UNITED STATES PATENT OFFICE CERTIFICATION OF CORRECTION Patent No. 2,979,518 April 11 1961 David Adriaan van Dorp et a1.

It is hereby certified that error appears in the above numbered patent requiring correction and 'that the said Letters Patent should read'es corrected below.

Column 2 line 35, for "btuyric" read butyric line 51, for "esteterification" read esterification column 4, line 32, for "3ket0ne" read 3-ketocolumn 5 line 26, strike out ".Foundz'7; column 6, line 58 for "71;" read 17a column 7, line 65, for "l g. read 18 g. line 65, for "1 m1." read 150 ml,

Signed and sealed this 29th'day of August 1961.

(SEAL) Attest:

ERNEST W. SWIDER DAVID L. LADD Attesting Officer Commissioner of Patents 

1. A COMPOUND SELECTED FROM GROUP CONSISTING OF OESTRADIOL-3,17-DI-ALANINATE AND THE ACID ADDITION SALTS THEREOF.
 3. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF CORTISONE-21-ALANINATE AND THE ACID ADDITION SALTS THEREOF. 